mRNA technology may have potential to repair, regenerate cardiac tissue

If this can create angiogenesis our stroke researchers should be jumping all over it for applications in the brain. But NOTHING WILL OCCUR. We have NO stroke leadership and NO stroke strategy.

mRNA technology may have potential to repair, regenerate cardiac tissue

By Erik Swain

By Regina Fritsche Danielson, PhD

By Ruchira Glaser, MD, MS

Source/Disclosures

  Tissue regeneration has been achieved in certain parts of the body, but not the heart. That could change if a novel therapeutic derived from messenger RNA succeeds.

AstraZeneca and Moderna are developing AZD8601, a locally administered messenger RNA (mRNA) therapy that encodes for vascular endothelial growth factor A. In preclinical and phase 1 studies, it demonstrated the potential for regenerative angiogenesis. It is now being evaluated for its ability to generate new cardiomyocytes and improve parameters in patients with conditions such as CVD and HF.

Ruchira Glaser, MD, MS, senior vice president and therapeutic area head of rare disease, autoimmune and cardiovascular at Moderna.

The phase 2 EPICCURE randomized trial, presented in November at the American Heart Association Scientific Sessions, was designed to evaluate the safety and tolerability of AZD8601. It included 11 patients with HF undergoing CABG, seven of whom were assigned an AZD8601 injection during their procedure and four of whom were assigned a placebo injection.

At 6 months, the agent was safe and well tolerated, and all seven patients in the AZD8601 group had N-terminal pro-B-type natriuretic peptide levels below the threshold for HF, compared with one of the four patients in the placebo group, whereas left ventricular ejection fraction and quality of life metrics also favored the AZD8601 group.

Regina Fritsche Danielson

The trial was led by Regina Fritsche Danielson, PhD, senior vice president and global head of research and early development, cardiovascular, renal and metabolic diseases at AstraZeneca, and Ruchira Glaser, MD, MS, senior vice president and therapeutic area head of rare disease, autoimmune and cardiovascular at Moderna.

Healio spoke to Danielson and Glaser about the effect of mRNA on cardiac tissues, how AZD8601 works, the implications of the EPICCURE trial, the next steps in development and the trial being the first female-led study of an mRNA therapy.

Healio: How can mRNA be used to repair or regenerate cardiac tissues?

Danielson: VEGF is a well-known angiogenic factor; its biological method of action is primarily to induce the formation of new blood vessels from existing vessels. The secondary role, which has been discovered more recently, is that VEGF can also activate epicardial stem cells located on the surface of the heart, and induce differentiation of those cells into cardiomyocytes and endothelial cells. Those endothelial cells can also contribute to new vascularization, but also, potentially new cardiomyocytes could contribute to cardiac contractility. That is a hypothesis that still needs to be truly proven, but there are some animal data supporting that differentiation of stem cells. Right now, the most validated function of VEGF is the formation of new vessels.

Healio: What is the potential for AZD8601 in the cardiology space?

Danielson: We see this as a unique molecule to repair a damaged heart; primarily a heart that is ischemic and does not have enough oxygen supply. This would typically be in patients after MI, when the heart loses function of vessels and gets scar tissue, which leads to poor contraction. In those patients, we see a great opportunity to modify the disease in terms of making small vessels. There is also perhaps a use in patients with HF or severe CAD requiring bypass surgery; that is not always sufficient because some of these patients have microvascular disease that cannot be cleared by a bypass. We believe VEGF is a treatment that can be added during CABG to support the formation of new vessels and reperfusion of the whole cardiac muscle to improve function.

Healio: Why is VEGF-A production important for certain patients with CV conditions?

Glaser: In addition to what Dr. Danielson just said, patients who have areas of the heart that are still alive but the blood flow is not good enough are the ones who could benefit from something which promotes blood vessel growth by giving the mRNA. The mRNA is just a message to the cell to produce the protein that it would normally produce under certain conditions. Patients who have MI or have CAD with lots of blockages and have heart muscle dysfunction as a result are the ones who could benefit.

Healio: Why did you undertake the EPICCURE study and what did you hope to learn from it?

Danielson: EPICCURE was primarily a safety and tolerability study. It was a feasibility study to help us understand how we could deliver mRNA into the heart in the best way. The study included patients who had open-chest CABG surgery so we could access the heart easily and administer the mRNA in a safe way. We also had a placebo group, because we wanted to understand the effect of injecting the drug itself vs. injecting a saline solution. We learned that this was safe and tolerable. We also learned we could easily administer the injections in a short time period, so we did not have to have the patient on the surgical table for much longer. We also explored the efficacy of the therapy. The results give us the excitement to continue the program forward with larger studies. This was a small study, with seven patients who were treated with the mRNA and four who received placebo in the form of saline solution.

Healio: What were the most important findings from EPICCURE?

Danielson: We demonstrated that the therapy is safe and tolerable. This is a procedure that is easy to carry out in any cardiology lab via direct injection into the heart. We are exploring other means of access, such as using percutaneous catheters, as a way to bring the therapy to more patients in the future if this is successful. We were also able to look at biomarker data, echocardiography data, etc to see that the therapy mimicked what we saw in preclinical studies. It is always exciting when that happens, because it shows this could potentially make a big difference for these patients.

Glaser: We have to be careful not to overinterpret the results, but it is exciting to see that all the traditional markers of HF trended in the right direction. The biomarker levels returned to normal in all treated patients and only one placebo patient. The echocardiogram looked like heart muscle function improved more in the treated patients. The quality of life questionnaire also indicated more improvement in the treated patients. While this is a small study, those positive signs that confirm that we might be seeing what we hypothesized we would see encourage us to go to the next step.

Healio: What are the next steps in the development of AZD8601?

Danielson: We want to move into a bigger study so we can assess the efficacy of the drug and continue to assess the safety and tolerability. And to move into patients with severe HF who are not responding enough to standard of care and may need CABG, or in the future may not be eligible for CABG and have no option and may need a transplant. We are going to start with those severe patients and are working on designing a larger trial with enough patients to make firm conclusions on efficacy in terms of cardiac function, using echocardiography and traditional biomarkers used in HF trials such as NT-proBNP, and also assessing the well-being of patients using Kansas City Cardiomyopathy Questionnaires, because we believe that based on increasing oxygen supply to the heart, making the heart pump better will make patients feel better, which is important for these patients who have poor quality of life without treatment.

Healio: Are there any other plans to apply mRNA technology to CV problems?

Danielson: We are working on many research activities using different modalities, one of which is the mRNA modality. We have a lot of hope for mRNA when it comes to cardiac disease. This is based on the finding that the uptake of mRNA in cardiac tissue can produce the NT-proBNP level that we would like it to produce. This is unique to the heart tissue. We are looking at other factors that can produce cardiomyocyte proliferation such as transdifferentiation of fibroblasts into cardiomyocytes. We at AstraZeneca are primarily focusing on cardiac repair and regeneration at the moment.

Glaser: At Moderna, we are also looking at more systemic applications for CVD and HF. We believe this is the beginning for regenerative therapeutics, including in CVD, which remains an area of extremely high unmet need globally. We are pursuing additional pathways where mRNA may have an advantage over traditional therapies.

Healio: Much has been written about the underrepresentation of women in the cardiology field. What does having a female-led CV mRNA study for the first time mean for the field?

Glaser: That is an interesting question. My background is in interventional cardiology, which has a small number of women in it. It is a fabulous additional theme that a diverse group of talent including women can bring amazing, innovative science to the field. It takes an entire team to do that. Hopefully, the leadership piece inspires other women, including younger students, to pursue a STEM career, so we can attract the best talent to treat diseases, a rewarding and important mission.

Danielson: If we can be role models and inspire younger women to give them confidence that they can become key players in their field, whether it is cardiology or anywhere in science, that is great.

Healio: Is there anything else you would like to mention?

Danielson: We are excited about these efforts, even though it is too early to promise anything yet. We are figuring out how we can regenerate tissue in the future, capitalizing on new technologies like mRNA, gene therapy, cell therapy and others to make this a reality. Our line of study could make a big difference to patients and translate into real benefits.

Glaser: We are happy we had the opportunity to collaborate with AstraZeneca because this is an area where people wanted to make progress for a long time, but we just did not have the right tools. The promise that mRNA could do something transformative for HF, which is so prevalent and devastating, is inspiring. We are anxious to do more studies to see if the promise holds true.

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Reference:

• Danielson RF, et al. Poster Presentations. Presented at: American Heart Association Scientific Sessions; Nov. 13-15, 2021 (virtual meeting).

 

hsa-miR-518-5p/hsa-miR-3135b Regulates the REL/SOD2 Pathway in Ischemic Cerebral Infarction

You described something but DID NOTHING TO SOLVE STROKE FOR SURVIVORS. Survivors are the reason for stroke research, didn't your mentors and senior researchers tell you that?

hsa-miR-518-5p/hsa-miR-3135b Regulates the REL/SOD2 Pathway in Ischemic Cerebral Infarction

Boyan Zhao and Xiaofan Jiang^*

• Department of Neurosurgery, Xijing Hospital, Fourth Military Medical University, Xi'an, China

Objectives: Ischemic cerebral infarction (ICI) is a fatal neurovascular disorder. A bioinformatics approach based on single-cell and bulk RNA-seq analyses was applied to investigate the pathways and genes involved in ICI and study the expression profile of these genes.

Methods: First, the aberrantly regulated “small-molecule ribonucleic acids” [microRNA (miRNAs)] and messenger RNAs (mRNAs) were analyzed using transcriptome data from the ischemic brain infarction dataset of the Gene Expression Omnibus (GEO) database. In mouse cerebrovascular monocytes, the single-cell regulatory network inference and clustering (SCENIC) workflow was used to identify key transcription factors (TFs). Then, the two miRNA-TF-mRNA interaction networks were constructed. Moreover, the molecular complex detection (MCODE) extracted the core sub-networks and identified the important TFs within these sub-networks. Finally, whole blood samples were collected for validation of the expression of critical molecules in ICI.

Results: We identified four cell types and 266 regulons in mouse cerebrovascular monocytes using SCENIC analysis. Moreover, 112 differently expressed miRNAs and 3,780 differentially expressed mRNAs were identified. We discovered potential biomarkers in ICI by building a miRNA-TF-mRNA interaction network. The hsa-miR-518-5p/hsa-miR-3135b/REL/SOD2 was found to play a potential role in ICI progression. The expression of REL and superoxide dismutase 2 (SOD2) was significantly elevated in the ICI group in the clinical cohort (P < 0.05). Furthermore, a REL expression was elevated in endothelial cells and fibroblasts at the single-cell level, indicating that REL is a cell-specific regulon. Functional enrichment analyses revealed that REL is primarily engaged in neurotransmitter activity and oxidative phosphorylation.

Conclusions: Our research uncovered novel biomarkers for ICI of neurovascular disease. The hsa-miR-518-5p/hsa-miR-3135b may regulate the REL/SOD2 pathway in ICI progression.

Introduction

Stroke is a common and fatal neurovascular disease that has high morbidity and mortality rates worldwide, accounting for ~17 million new cases annually (1–5). Ischemic stroke (IS) accounts for 80% of all stroke cases (6). Multiple emboli blocking the intracerebral arteries result in irreversible functional deficiencies in local brain tissue, eventually leading to ischemia and hypoxic necrosis (7, 8). Ischemic cerebral infarction (ICI) is a fatal neurovascular disorder (9). Due to its complexity, the molecular pathways underlying the development of ICI are not well-known at the transcriptome level. Exploring the regulatory network of signaling pathways is critical to understanding the mechanism by which ICI develops and to developing effective strategies for preventing and treating ICI.

Single-cell RNA sequencing (scRNA-seq) is a technique for obtaining whole-transcriptome expression profiles at the single-cell level. It is based on the amplification of microscopic whole-transcriptome RNA from isolated individual cells and subsequent high-throughput sequencing to elucidate the molecular regulatory mechanisms underlying specific biological processes and disease pathogenesis (10). In recent years, the scRNA-seq has gradually gained traction in the disciplines of oncology, microbiology, and neuroscience (11–13). In addition to studying changes in gene expression patterns at the population level, the scRNA-seq can be used to study single-cell gene expression, thus, resolving any bias arising due to cellular heterogeneity. Therefore, scRNA-seq is particularly well-suited for studying highly heterogeneous cell populations, such as neural cells (14, 15). Using scRNA-seq, Gate et al. showed that the T-cell receptor (TCR) signaling pathway is activated in CD8+ terminally differentiated effector cells (TEMRA) in the cerebrospinal fluid of patients with Alzheimer's disease, indicating that these cells contribute to the onset of neurological symptoms through their cytotoxic role (16). Vanlandewijck et al. used scRNA-seq to conduct a transcriptional investigation of the constituent cell types of the cerebral vasculature and discovered that endothelial cells, pericytes, and fibroblasts are implicated in the formation of neurovascular disease lesions. In this study, we attempted to address the dearth of molecular studies on cerebrovascular cell types and to establish the groundwork for future research on the molecular pathways underlying cerebrovascular disorders (17). The single-cell regulatory network infeAbegail Floresrence and clustering (SCENIC) is a computational approach for identifying cell states and constructing gene regulatory networks from scRNA-seq data (18). The SCENIC can be used to identify critical transcription factors (TFs) involved in the pathophysiology of ICI.

Apart from single-cell technology, various bioinformatics techniques have emerged as important tools to study complex biological phenomena. The “small-molecule ribonucleic acids” [microRNA (miRNAs)] are a class of non-coding, endogenous single-stranded RNA molecules composed of 20–24 nucleotides that regulate the expression of target genes in various physiological and pathological processes (19). The miRNAs serve as molecular markers for early diagnosis and prognosis, as well as therapeutic targets for ICI (20–23). The miR-PC-5P-12969 inhibits the production of amyloid and promotes IS (24). In acute ICI, the serum miR-124 and other miRs are inhibited, resulting in neuroinflammation and brain damage (25). Additionally, miRNAs can contribute to ICI by modulating TFs. Atherosclerosis (AS) has a significant role in the pathophysiology of ICI (26). Li et al. showed that miR-NA-663 governs the phenotypic metamorphosis of human vascular smooth muscle cells by adversely regulating the expression of its downstream TF, Jun B (27). The miRNA-23b can reduce vascular smooth muscle cell proliferation and migration, and the TF FoxO4 may be a direct target of miRNA-23b (28). Thus, the miRNAs play a critical role in the development of AS by controlling the proliferation, differentiation, and function of vascular smooth muscle cells via TF regulation, potentially altering the course of ICI. Therefore, single-cell sequencing is critical for identifying miRNA-TF-gene regulatory networks involved in ICI progression, which may reveal novel gene targets and molecular markers for ICI diagnosis and therapy.

In this study, we analyzed bulk RNA-seq, as well as scRNA-seq data, to identify a miRNA-TF-mRNA regulatory network that may be vital to ICI progression. Our research aimed to uncover novel biomarkers for ICI of neurovascular disease. The findings of this study may provide new avenues for the prevention and treatment of IS, as well as strategies to improve patient outcomes.

More at link.

 

A master regulatory network restoring brain glutamate homeostasis is coordinately activated in stroke

Somewhere in these 29 pages something might be useful. Which our great stroke association would analyze and distribute worldwide. Because right now your doctor and stroke hospital are not updating their stroke recovery protocols at all, they don't have any. 

Do you prefer your doctor, hospital and board of director's incompetence NOT KNOWING? OR NOT DOING?

A master regulatory network restoring brain glutamate homeostasis is coordinately activated in stroke

  Mariko Kobayashi1,3, Corey Anderson2, Corinne Benakis2, Michael J. Moore1, Aldo Mele1, John J. Fak1, Christopher Y. Park1, Ping Zhou2, Josef Anrather2, Costantino Iadecola2, Robert B. Darnell1,3. 1Laboratory of Molecular Neuro-Oncology and Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA. 2Fell Family Brain and Mind Research Institute, Weill Cornell Medicine, 407 East 61st Street, New York, NY, 10065, USA. 3Correspondence: mkobayashi@rockefeller.edu, darnelr@rockefeller.edu

ABSTRACT
Altered miRNA expression in various disease states have been identified, but their global targets contributing to the collective regulatory power to promote or attenuate pathology remains poorly defined. Here we applied a combination of hi-throughput RNA profiling techniques, including AGO CLIP, miRNAseq, RNAseq and ribosomal profiling, to develop an unbiased and comprehensive view of miRNA:mRNA functional interactions following ischemia/reperfusion (IR) injury in the mouse brain. Upon acute I/R insult miR-29 family members were most prominently lost, with corresponding de-regulation of their global target sites. This leads to a dynamic, cascading mode of miR-29 target transcript activation, orchestrated by an initial translational activation and subsequent increase in target mRNA levels. Unexpectedly, activated genes include factors essential for glutamate signaling and reuptake, indicating a fundamental role for this regulatory network in modulating criticalendogenous neuroprotective programs to restore brain homeostasis. We integrated this data with human brain AGO CLIP profiles to infer target site variants that determine miRNA binding and to explore the role of non-coding site polymorphisms in stroke. Together these results establish a new strategy for understanding RNA regulatory networks in complex neurological disease.

Interactive effects of cell therapy and rehabilitation realize the full potential of neurogenesis in brain injury model

If I'm reading this correctly, any stem cell transplantation should be accompanied by exercise.
http://www.sciencedirect.com/science/article/pii/S0304394013008252

• Takeshi Imura^a,
• Masaya Matsumoto^a,
• Takahiro Fukazawa^a,
• Elham Khalesi^a,
• Yanan Sun^a,
• Masaaki Takeda^b,
• Hiroyuki Uwatoko^a,
• Kyosuke Nakata^a,
• Keiji Tanimoto^c,
• Teruyuki Kajiume^d,
• Yumi Kawahara^e,
• Louis Yuge^{a, e, ,}

• ^a Division of Bio-Environmental Adaptation Sciences, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan
• ^b Department of Neurosurgery, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan
• ^c Department of Radiation Medicine, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan
• ^d Department of Pediatrics, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan
• ^e Space Bio-Laboratories Co., Ltd, Hiroshima, Japan

---

Highlights

•

Combined cell therapy and rehabilitation enhances functional and electrophysiological recovery in brain-injured mice.

•

Rehabilitation may facilitate the differentiation of transplanted neural stem/progenitor cells into functional neurons.

•

mRNAs involved in neural plasticity (BDNF, GAP43) were up-regulated by combined rehabilitation and cell transplantation

---

Abstract

The therapeutic effect of rehabilitation after cell therapy for brain injury remains unclear. Here, we report the neural stem/progenitor cells transplantation into a brain injury mouse model followed by treadmill exercise training. Among all experimental groups, mice that underwent transplantation and treadmill exercise demonstrated significant functional motor and electrophysiological improvement. Transplanted cells at the brain injury site were observed and differentiated into neurons and astrocytes. Transplanted cells significantly differentiated into neurons in the mice that underwent transplantation and treadmill exercise compared with those treated with only transplantation. Furthermore, the expression of brain-derived neurotrophic factor and growth-associated protein 43 mRNAs were significantly up-regulated in the mice that underwent transplantation and treadmill exercise than in those in other experimental groups during the early recovery stage. These results suggest that rehabilitation after neural stem/progenitor cell transplantation enhances neurogenesis and promotes the recovery of motor function in brain injury model mice

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Caterpillar fungi may benefit asthma patients

Its got anti-inflammatory properties and inflamation in the brain is a problem. Get your researcher involved.

When I was in Bhutan you could buy this, I didn't buy any. Ask your doctor what research they are doing on this.

All these benefits needing scientific proof. #6 seems really important.

Caterpillar fungi may benefit asthma patients

Research shows that cordycepin, extracted from a group of rare parasitic caterpillar fungi of the genus Cordyceps, has unusual anti-inflammatory properties that may make it a good candidate for treating conditions such as asthma and rheumatoid arthritis.
The investigators, led by Cornelia de Moor (University of Nottingham, UK), found that cordycepin, which is similar in structure to adenosine, inhibits the stimulation of inflammatory messenger RNAs (mRNAs) by cytokines secreted by smooth muscle cells in the human airway, but does not affect the expression of "housekeeping" mRNAs.
"We have shown that cordycepin reduces the expression of inflammatory genes in airway smooth muscle cells by acting on the final step in the synthesis of their messenger RNAs (mRNAs) which carry the chemical blueprint for the synthesis of proteins. This process is called polyadenylation," said de Moor in a press statement.
"However, it is a surprise that cordycepin does not affect the synthesis of mRNAs from other genes, because nearly all mRNAs require polyadenylation," she said.
Cordyceps fungi live on hibernating caterpillars in the Tibetan mountains and have been a highly sought after component of Chinese medicine for many years. Studies have suggested the fungi could be used to treat a variety of conditions including cancer, stroke, kidney disease, and inflammatory lung disease, but scientific evidence regarding the mechanism of action of cordycepin was unclear until now.

In Vitro Modelling of Cortical Neurogenesis by Sequential Induction of Human Umbilical Cord Blood Stem Cells

I hope someone understands this. 

In Vitro Modelling of Cortical Neurogenesis by Sequential Induction of Human Umbilical Cord Blood Stem Cells

Résumé / Abstract

Neurogenesis of excitatory neurons in the developing human cerebral neocortex is a complex and dynamic set of processes and the exact mechanisms controlling the specification of human neocortical neuron subtypes are poorly understood due to lack of relevant cell models available. It has been shown that the transcription factors Pax6, Tbr2 and Tbr1, which are sequentially expressed in the rodent neocortex, regulate and define corticogenesis of glutamatergic neocortical neurons. In humans the homologues of these genes are generally expressed in a similar pattern, but with some differences. In this study, we used purified human umbilical cord blood stem cells, expressing pluripotency marker genes (OCT4, SOX2 and NANOG), to model human neocortical neurogenesis in vitro. We analyzed the expression patterns of PAX6, TBR2 and TBR1, at both protein and mRNA levels, throughout the 24 days of a sequential neuronal induction protocol. Their expression patterns correlated with those found in the developing human neocortex where they define different developmental stages of neocortical neurons. The derived cord blood neuron-like cells expressed a number of neuronal markers. They also expressed components of glutamatergic neurotransmission including glutamate receptor subunits and transporters, and generated calcium influxes upon stimulation with glutamate. Thus we have demonstrated that it is possible to model neocortical neurogenesis using cord blood stem cells in vitro. This may allow detailed analysis of the molecular mechanisms regulating neocortical neuronal specification, thus aiding the development of potential therapeutic tools for diseases and injuries of the cerebral cortex.